ET(A) and ET(B) specific ligands synergistically antagonize endothelin-1 binding to an atypical endothelin receptor in primary rat astrocytes.
Using a whole-cell binding procedure with long incubations at low temperature and subsequent acid stripping, we have characterized an atypical endothelin (ET) receptor in primary rat cortical astrocyte cultures. We found the following: (a) no competition for 125I-ET-1 binding by the ET(A) antagonists BQ-123 and LU 135252 or the ET(B) agonist IRL 1620; (b) weak competition by the ET(B) antagonist BQ-788 and by the predominant ET(B) ligand ET-3; (c) potent synergistic competition of ET(A) and ET(B) ligands in combination for 125I-ET-1 binding; (d) potent competition of ET-1 with any of the radioligands used, 125I-ET-1, 125I-IRL 1620, and [3H]BQ-123; (e) lack of competition of IRL 1620 and BQ-123 with the respective other radioligand; (f) shifting of the amount of acid-strippable 125I-ET-1 binding from 20 to 80% by ET(B) ligands and to 4% by ET(A) ligands; and (g) as a control, typical ET(A) and ET(B) binding characteristics of the RAT-1 fibroblast and the U373MG astrocytoma cell line, respectively, under our assay conditions. The unusual binding properties of astrocytic ET receptors described in this study appear to be the result of several binding sites in the receptor for different ET ligands or ligand epitopes.
Jensen
N
N
Department of Neurology, Georg-August-University, and Max-Planck-Institute for Experimental Medicine, Göttingen, Germany.
Hasselblatt
M
M
Sirén
A L
AL
Schilling
L
L
Schmidt
M
M
Ehrenreich
H
H
D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000378:metabolism*; D001665:Binding Sites; D001667:Binding, Competitive; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000378:metabolism*; D065128:Endothelin Receptor Antagonists; D019332:Endothelin-1 / Q000378:metabolism*; D016232:Endothelins / Q000378:metabolism / Q000494:pharmacology*; D007700:Kinetics; D008024:Ligands; D008958:Models, Molecular; D010446:Peptide Fragments / Q000378:metabolism / Q000494:pharmacology*; D010456:Peptides, Cyclic / Q000378:metabolism / Q000494:pharmacology*; D011487:Protein Conformation; D011869:Radioligand Assay; D051381:Rats; D017208:Rats, Wistar; D044022:Receptor, Endothelin A; D044023:Receptor, Endothelin B; D017466:Receptors, Endothelin / Q000145:classification / Q000378:metabolism*
D065128:Endothelin Receptor Antagonists; D019332:Endothelin-1; D016232:Endothelins; D008024:Ligands; D010446:Peptide Fragments; D010456:Peptides, Cyclic; D044022:Receptor, Endothelin A; D044023:Receptor, Endothelin B; D017466:Receptors, Endothelin; C074308:sovateltide; C072247:cyclo(Trp-Asp-Pro-Val-Leu)